本文選題：交感神經(jīng) + 心房顫動(dòng)　； 參考：《廣西醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的本課題旨在通過(guò)建立擬交感心房顫動(dòng)模型,研究心房縫隙連接蛋白43(connexin 43,Cx43)表達(dá)水平及分布的變化,從而探討交感性房顫發(fā)生與維持的可能的內(nèi)在機(jī)制。方法建立交感性心房顫動(dòng)模型,健康幼犬15只,雌雄不分,將其隨機(jī)分成3組,每組5只,分別為:1.對(duì)照組(Control組),將實(shí)驗(yàn)犬心臟取出后,快速連接至langendorff心臟離體灌流儀,予臺(tái)氏液灌流,持續(xù)約1h�？焖傩姆科鸩M(rapid atrial pacing,RAP組),灌流方法同Control組,僅在灌流時(shí)用電生理刺激儀以800次/min頻率起搏30s,共30次。異丙腎上腺素灌流組+快速心房起搏組(isoprenaline+rapid atrial pacing,ISO+RAP組),以含0.1umol/L異丙腎上腺素(isoprenaline,ISO)的改良臺(tái)氏液灌流心臟,余同RAP組。(1)在離體灌流狀態(tài)下,通過(guò)快速心房起搏構(gòu)建擬交感性房顫模型。各組分別測(cè)量心房有效不應(yīng)期(atrial effective refractory period,AERP)和記錄各組的房顫誘發(fā)率的情況。實(shí)驗(yàn)結(jié)束后,取左心房組織固定,通過(guò)免疫組化檢測(cè)心房神經(jīng)生長(zhǎng)因子(NGF)和酪氨酸羥化酶(TH)的表達(dá)和分布變化。觀察擬交感效應(yīng)對(duì)快速起搏離體心臟的影響。(2)取部分左心房組織,采用western blotting方法檢測(cè)各組心房肌中總Cx43的含量及磷酸化Cx43的含量,Q-PCR檢測(cè)Cx43mRNA表達(dá)量,免疫熒光(immunofluorescence,IF)檢測(cè)心房肌組織Cx43的分布變化,凋亡試劑盒檢測(cè)心房肌組織的細(xì)胞凋亡率,透射電鏡檢測(cè)心房肌線粒體形態(tài)的變化,熒光比色法檢測(cè)心房肌線粒體ROS的生成量。結(jié)果(1)各組所測(cè)得的AERP中,Control組和RAP組無(wú)顯著差別(P0.05),ISO+RAP組AERP則顯著縮短(P0.05);ISO+RAP組可成功誘發(fā)房顫,而Control組及RAP組均無(wú)法誘發(fā)房顫;與Control組比較,RAP組及ISO+RAP組NGF和TH的含量及分布均增多,ISO+RAP組高于RAP組。(2)與Control組比較,RAP組及ISO+RAP組Cx43及磷酸化Cx43含量呈逐漸降低趨勢(shì)(P0.05);與Control組比較,RAP組Cx43分布呈明顯側(cè)向化,線粒體輕度腫脹,基質(zhì)完整,ISO+RAP組Cx43呈點(diǎn)狀散在分布,線粒體明顯腫脹,部分基質(zhì)透明;ISO+RAP組細(xì)胞凋亡指數(shù)(apoptosis index,AI)及線粒體ROS生成量均顯著高于RAP組及Control組(P0.05),而RAP組則高于Control組(P0.05)。結(jié)論1.通過(guò)異丙腎上腺素灌流模擬交感神經(jīng)興奮效應(yīng),可以顯著縮短心房肌的動(dòng)作電位時(shí)程,表現(xiàn)為AERP時(shí)程縮短。2.單純的高頻心房起搏在本實(shí)驗(yàn)持續(xù)的時(shí)間內(nèi)無(wú)法誘發(fā)房顫,而在擬交感效應(yīng)下則可以成功誘發(fā),成功構(gòu)建擬交感性房顫模型。3.在交感性心房顫動(dòng)中,交感神經(jīng)可能通過(guò)氧化應(yīng)激效應(yīng),引起Cx43的重構(gòu)與下調(diào),從而介導(dǎo)心房顫動(dòng)的發(fā)生。
[Abstract]:Objective to study the changes of the expression and distribution of atrial gap junction protein (43(connexin 43) by establishing a sympathetic atrial fibrillation model, and to explore the possible mechanism of occurrence and maintenance of sympathetic atrial fibrillation. Methods A model of sympathetic atrial fibrillation was established. Fifteen healthy puppies were randomly divided into 3 groups, 5 in each group: 1: 1. After the heart was removed from the experimental dog, the control group was quickly connected to the langendorff heart perfusion apparatus in vitro and perfused with Tyrtle fluid for about 1 hour. In the rapid atrial pacing group, the perfusion method was the same as that in the control group. Isoprenaline rapid atrial pacing group (IPA) was perfused with modified Tyrna solution containing 0.1 umol / L isoproterenol (isoproterenol) and isoprenaline isoprenalineiso (ISO), and the rest was the same as that in rap group under isolated perfusion. A sympathetic atrial fibrillation model was constructed by rapid atrial pacing. Atrial effective refractory AERP (AERP) and atrial fibrillation evoked rate were measured in each group. The expression and distribution of nerve growth factor NGF and tyrosine hydroxylase were detected by immunohistochemistry. To observe the effect of pseudo sympathetic effect on the isolated heart of rapid pacing, some left atrial tissues were taken. The contents of total Cx43 and phosphorylated Cx43 were detected by western blotting method and the expression of Cx43 mRNA was detected by Q-PCR. The distribution of Cx43 in atrial myocytes was detected by immunofluorescence fluorescence assay, apoptosis rate in atrial muscle tissue was detected by apoptosis kit, mitochondrial morphology of atrial muscle was detected by transmission electron microscope, Ros production in mitochondria of atrial muscle was detected by fluorescence colorimetry. Results 1) there was no significant difference in AERP between control group and rap group. AERP in ISO rap group was significantly shorter than that in ISO rap group, but atrial fibrillation could not be induced in control group and rap group. The contents and distribution of NGF and th in rap group and ISO rap group were higher than those in rap group.) compared with control group, the contents of Cx43 and phosphorylated Cx43 in rap group and ISO rap group decreased gradually (P 0.05), and the distribution of Cx43 in rap group was more lateral than that in control group. The mitochondria were slightly swollen, the distribution of Cx43 in ISO rap group was dotted, the mitochondria was obviously swollen, and the apoptosis index (AI) and Ros production of mitochondria in ISO rap group were significantly higher than those in rap group and Control group, while that in rap group was higher than that in Control group. Conclusion 1. By using isoproterenol perfusion to simulate sympathetic excitatory effect, the action potential duration of atrial muscle can be significantly shortened, as shown by the shortening of AERP duration by .2. High frequency atrial pacing alone could not induce atrial fibrillation during the duration of the experiment, but could be induced successfully under the pseudo sympathetic effect, and successfully constructed the pseudo sympathetic atrial fibrillation model. 3. In the process of sympathetic atrial fibrillation, sympathetic nerve may induce the remodeling and down-regulation of Cx43 through oxidative stress, thus mediating the occurrence of atrial fibrillation.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R541.75;R-332

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